The Albany High Voltage Electron Microscope (HVEM) Biotechnological Resource currently serves a large number of researchers in the Northeastern U.S. The main features of the Resource include the 11.2 MVAEI HVEM and STERECON, a unique computer-based system for Resource into a comprehensive Biological Microscopy and Image Reconstruction Resource to better serve the rapidly changing and increasingly sophisticated needs for structural analysis within the Biomedical community. This goal will be accomplished by improving existing facilities and by incorporating into the Resource new capabilities developed at the applicant institution over the past 5 years. These include HVEM tomography and confocal light microscopy (LM), which have been developed with NIH Resource-Related grants. Other facilities to be added (funded by other awards) include a high-resolution, 120-kV Philips EM 420 equipped for cryo-operation, a video-enhanced LM system, and the complete SPIDER image processing system (developed in-house), which has been installed on a new VAX workstation network. This proposed Resource will be unique in that it will allow users to conduct correlative LM and EM studies at the highest possible resolution and to obtain 3D ultrastructural data encompassing a wide range of cellular organization from macromolecules and organelles to whole cells and tissues. The specific aims of this application are first to efficiently integrate the various facilities described above into a single, user-friendly Resource and then to develop specific aspects of these tools which will further enhance the technological capabilities of the Resource. These aspects include: 1) the development of cryofixation techniques for correlative LM/EM studies of cells followed by video-enhanced LM, 2) further development of confocal LM to increase its usefulness for 3D LM and correlative LM/EM studies, 3) upgrading the existing HVEM tilt stages and developing new stages for high resolution tomographic and STERECON applications, and 4) the development of image restoration procedures for use in tomographic and other 3D reconstruction techniques. A large and growing number of NIH-funded collaborative and service projects currently utilize the facilities within the existing Resource. This expansion of the user community can be expected to continue when the new technological capabilities, already in place and in demand by external investigators, are integrated into the Resource.